The unique functions of the liver require detailed study, since hepatopathology is a serious veterinary problem, which also negatively affects the productivity of farm animals. It is possible to clarify the pathogenetic mechanisms and its development using artificial modeling of hepatopathology and determine the therapeutic efficacy of hepatoprotective drugs, especially based on raw materials of animal origin. The aim of the study was to determine the effect of the use of milk phospholipids in the composition of the bioadditive “FLP-MD” on the secretion of bile acids by the liver in artificial modeling of fatty hepatosis in rats. To reproduce hepatopathology, laboratory rats were administered intragastrically with a 4% solution of tetracycline hydrochloride at a dose of 0.25 g/kg of body weight for seven days and the bioadditive was used at a dose of 13.5 mg/kg of body weight for nine days. Bile samples were collected from rats by conducting acute experiments. Six fractions of conjugated bile acids were determined in bile samples by thin-layer chromatography. It was found that in laboratory rats with experimental fatty hepatosis, the processes of biotransformation of primary and secondary cholates by conjugation with taurine were inhibited. In particular, a decrease in the concentration of taurocholic acid in the bile of sick animals by 20.5%-38.1% (P < 0.01), and of the complex of taurochenodeoxycholic and taurodeoxycholic acids by 21.8%-25.7% (P < 0.05) was recorded. In the case of using the bioadditive “FLP-MD” in rats with experimental fatty hepatosis, the concentration of taurocholic, taurochenodeoxycholic and taurodeoxycholic acids in bile significantly increased. The concentration of glycoconjugated bile acids and free cholates corresponded to their level in the control. The use of the bioadditive “FLP-MD” based on milk phospholipids in experimental fatty hepatosis eliminated the negative impact of the antibiotic in a toxic dose on the processes of biotransformation and the formation of cholates. This allows to recommend the bioadditive “FLP-MD” based on milk phospholipids as a hepatoprotective agent in the case of the use of antimicrobial drugs in animals
bile acids; cholesterol; bioadditive “FLP-MD”; tetracycline hydrochloride; hepatocyte; hepatopathology
[1] Allameh, A., Niayesh-Mehr, R., Aliarab, A., Sebastiani, G., & Pantopoulos, K. (2023). Oxidative stress in liver pathophysiology and disease. Antioxidants (Basel), 12(9), article number 1653. doi: 10.3390/antiox12091653.
[2] Boyer, J.L., & Soroka, C.J. (2021). Bile formation and secretion: An update. Journal of Hepatology, 75(1), 190-201. doi: 10.1016/j.jhep.2021.02.011.
[3] Choudhuri, S., & Klaassen, C.D. (2022). Molecular regulation of bile acid homeostasis. Drug Metabolism and Disposition, 50(4), 425-455. doi: 10.1124/dmd.121.000643.
[4] de Bruijn, V.M.P., Wang, Z., Bakker, W., Zheng, W., Spee, B., & Bouwmeester, H. (2022). Hepatic bile acid synthesis and secretion: Comparison of in vitro methods. Toxicology Letters, 365, 46-60. doi: 10.1016/j.toxlet.2022.06.004.
[5] Deo, V., & Ranganathan, P. (2024). Statistical tools and packages for data collection, management, and analysis – a brief guide for health and biomedical researchers. Perspectives in Clinical Research, 15(4), 209-212. doi: 10.4103/picr.picr_160_24.
[6] European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes. (1986, March). Retrieved from https://rm.coe.int/168007a67b.
[7] Farooqui, N., Elhence, A., & Shalimar. (2022). A current understanding of bile acids in chronic liver disease. Journal of Clinical and Experimental Hepatology, 12(1), 155-173. doi: 10.1016/j.jceh.2021.08.017.
[8] Fleishman, J.S., & Kumar, S. (2024). Bile acid metabolism and signaling in health and disease: Molecular mechanisms and therapeutic targets. Signal Transduction and Targeted Therapy, 9(1), article number 97. doi: 10.1038/s41392-024-01811-6.
[9] Fu, Y., Guzior, D.V., Okros, M., Bridges, C., Rosset, S.L., González, C.T., Martin, C., Karunarathne, H., Watson, V.E., & Quinn, R.A. (2025). Balance between bile acid conjugation and hydrolysis activity can alter outcomes of gut inflammation. Nature Communications, 16(1), article number 3434. doi: 10.1038/s41467-025-58649-x.
[10] Fuchs, C.D., Simbrunner, B., Baumgartner, M., Campbell, C., Reiberger, T., & Trauner, M. (2025). Bile acid metabolism and signalling in liver disease. Journal of Hepatology, 82(1), 134-153. doi: 10.1016/j.jhep.2024.09.032.
[11] Guo, J., Chen, S., Zhang, Y., Liu, J., Jiang, L., Hu, L., Yao, K., Yu, Y., & Chen, X. (2024). Cholesterol metabolism: Physiological regulation and diseases. MedComm, 5(2), article number e476. doi: 10.1002/mco2.476.
[12] Ikeda, Y. (2020). Mechanism of taurohyodeoxycholate-induced biliary phospholipid efflux –understanding the function of the ABCB4 enhancer for developing therapeutic agents against bile salt-induced liver injury. Yakugaku Zasshi, 140(11), 1329-1334. doi: 10.1248/ yakushi.20-00156.
[13] Jia, W., Li, Y., Cheung, K.C.P., & Zheng, X. (2024). Bile acid signaling in the regulation of whole body metabolic and immunological homeostasis. Science China. Life Sciences, 67(5), 865-878. doi: 10.1007/s11427-023-2353-0.
[14] Korolova, D., Gryshchenko, V., Chernyshenko, T., Platonov, O., Hornytska, O., Chernyshenko, V., Klymenko, P., Reshetnik, Y., & Platonova, T. (2023). Blood coagulation factors and platelet response to drug-induced hepatitis and hepatosis in rats. Animal Models and Experimental Medicine, 6(1), 66-73. doi: 10.1002/ame2.12301.
[15] Law of Ukraine No. 3447 “On the Protection of Animals from Cruelty”. (2006, February). Retrieved from https://zakon.rada.gov.ua/laws/show/3447-15#Text.
[16] Liashevych, A.M., Lupaina, І.S., Davydovska, T.L., Tsymbalyuk, O.V., Oksentiuk, Y.R., & Makarchuk, M.Y. (2021). The effect of Corvitin on the content of bile acids in the liver of rats under conditions of chronic social stress. Regulatory Mechanisms in Biosystems, 12(3), 419-424. doi: 10.15421/022157.
[17] Lupaina, І., Liashevych, А., Reshetnik, Y., Veselsky, S., & Makarchuk, М. (2021). The effect of testosterone on the bile acid and bile lipid composition in rats. Scientific Reports of the National University of Life and Environmental Sciences of Ukraine, 17(5), 28-38. doi: 10.31548/ dopovidi2021.05.003.
[18] Melnychuk, D., Hryshchenko, V., & Litvinenko, O. (2009). Veterinary bioactive addidition of liposomal form and method of reparative therapy in hepatology. (Patent of Ukraine No. 86516). Retrieved from https://sis.nipo.gov.ua/uk/search/detail/422804/.
[19] Melnychuk, D.O., & Hryshchenko, V.A. (2014). Exchange of bile pigments under the action of ecopathogenic factors on organism. The Ukrainian Biochemical Journal, 86(5), article number 156.
[20] Miyazaki, T., Ueda, H., Ikegami, T., & Honda, A. (2023). Upregulation of taurine biosynthesis and bile acid conjugation with taurine through FXR in a mouse model with human-like bile acid composition. Metabolites, 13(7), article number 824. doi: 10.3390/metabo13070824.
[21] Nakamura, T., Masuda, A., Nakano, D., Amano, K., Sano, T., Nakano, M., & Kawaguchi, T. (2025). Pathogenic mechanisms of metabolic dysfunction-associated steatotic liver disease (MASLD)-associated hepatocellular carcinoma. Cells, 14(6), article number 428. doi: 10.3390/ cells14060428.
[22] Nemeth, K., Sterczer, Á., Kiss, D.S., Lányi, R.K., Hemző, V., Vámos, K., Bartha, T., Buzás, A., & Lányi, K. (2024). Determination of bile acids in canine biological samples: Diagnostic significance. Metabolites, 14(4), article number 178. doi: 10.3390/metabo14040178.
[23] Nikolajevic, N., Nikolajevic, M., Pantic, I., Korica, B., Kotseva, M., Alempijevic, T., Jevtic, D., Madrid, C.I., & Dumic, I. (2024). Drug-induced liver injury due to doxycycline: A case report and review of literature. Cureus, 16(5), article number e59687. doi: 10.7759/cureus.59687.
[24] Nimer, N., Choucair, I., Wang, Z., Nemet, I., Li, L., Gukasyan, J., Weeks, T.L., Alkhouri, N., Zein, N., Tang, W.H.W., Fischbach, M.A., Brown, J.M., Allayee, H., Dasarathy, S., Gogonea, V., & Hazen, S.L. (2021). Bile acids profile, histopathological indices and genetic variants for nonalcoholic fatty liver disease progression. Metabolism: Clinical and Experimental, 116, article number 154457. doi: 10.1016/j.metabol.2020.154457.
[25] Qi, Y., Ma, Y., & Duan, G. (2024) Pharmacological mechanisms of bile acids targeting the farnesoid X receptor. International Journal of Molecular Sciences, 25(24), article number 13656. doi: 10.3390/ijms252413656.
[26] Silva, C., Merim, S., Sevivas, R., Mota, J., & Leitão, A. (2023). Bismuth subcitrate, metronidazole and tetracycline – a rare cause of drug-induced liver injury. European Journal of Case ]Reports in Internal Medicine, 10(12), article number 004119. doi: 10.12890/2023_004119.
[27] Stellaard, F., & Lütjohann, D. (2021). Dynamics of the enterohepatic circulation of bile acids in healthy humans. American Journal of Physiology. Gastrointestinal and Liver Physiology, 321(1), G55-G66. doi: 10.1152/ajpgi.00476.2020.
[28] Taylor, K., et al. (2025) Perspective: How complex in vitro models are addressing the challenges of predicting drug-induced liver injury. Frontiers in Drug Discovery, 5, article number 1536756. doi: 10.3389/fddsv.2025.1536756.
[29] Tiwari, V., Shandily, S., Albert, J., Mishra, V., Dikkatwar, M., Singh, R., Sah, S.K., & Chand, S. (2025). Insights into medication-induced liver injury: Understanding and management strategies. Toxicology Reports, 14, article number 101976. doi: 10.1016/j.toxrep.2025.101976.
[30] Varma, S., Nathanson, J., Dowlatshahi, M., Del Portillo, A., Ramirez, I., & Garcia-Carrasquillo, R. (2021). Doxycycline-induced cholestatic liver injury. Clinical Journal of Gastroenterology, 14(5), 1503-1510. doi: 10.1007/s12328-021-01475-7.
[31] Ve[selskyi, S., Liaschenko, P., Kostenko, S., Horenko, Z., & Kurovska, L. (2001). The method of preparing samples of biofluids for determining the content of substances of a lipid nature. (Patent of Ukraine No. 33564). Retrieved from https://sis.nipo.gov.ua/uk/search/detail/342563/.
[32] Wei, C., Liu, Y., Jiang, A., & Wu, B. (2022). A pharmacovigilance study of the association between tetracyclines and hepatotoxicity based on Food and Drug Administration adverse event reporting system data. International Journal of Clinical Pharmacy, 44(3), 709-716. doi: 10.1007/s11096-022-01397-5.
[33] Wupperfeld, D., Fricker, G., Bois De Fer, B., & Popovic, B. (2024). Essential phospholipids impact cytokine secretion and alter lipid-metabolizing enzymes in human hepatocyte cell lines. Pharmacological Reports: PR, 76(3), 572-584. doi: 10.1007/s43440-024-00595-4.
[34] Wupperfeld, D., Fricker, G., Bois De Fer, B., Frank, L., Wehrle, A., & Popovic, B. (2022). Essential phospholipids decrease apoptosis and increase membrane transport in human hepatocyte cell lines. Lipids in Health and Disease, 21(1), article number 91. doi: 10.1186/s12944-022-01698-8.
[35] Yeo, X.Y., Tan, L.Y., Chae, W.R., Lee, D.Y., Lee, Y.A., Wuestefeld, T., & Jung, S. (2023). Liver’s influence on the brain through the action of bile acids. Frontiers in Neuroscience, 17, article number 1123967. doi: 10.3389/fnins.2023.1123967.
[36] Zhang, D., Zheng, J., Qiu, G., Niu, T., Gong, Y., & Cui, S. (2022). CCl4 inhibits the expressions of hepatic taurine biosynthetic enzymes and taurine synthesis in the progression of mouse liver fibrosis. Human & Experimental Toxicology, 41, article number 9603271221135033. doi: 10.1177/09603271221135033.